1. Cloning, sequencing, and recombinant production of Sin a 2, an allergenic 11S globulin from yellow mustard seeds 
Oscar Palomares, PhD, Andrea Vereda, MD, Javier Cuesta-Herranz, MD, PhD, Mayte Villalba, PhD, Rosalía Rodríguez, PhD 
Journal of Allergy and Clinical Immunology 
Volume 119, Issue 5, Pages (May 2007)
DOI: /j.jaci
Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2. Fig 1
Nucleotide sequence of cDNA encoding Sin a 2 subunit and deduced amino acid sequence. Cleavage site for signal peptide is indicated by an arrow head. ∗Potential cleavage site to render the chains of the mature protein. Peptides identified in the natural allergen13 are underlined. Numerals on the right side indicate amino acid position in the mature protein.
Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci )
Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3. Fig 2
Alignment of sequences of allergenic 11S globulins. Yellow mustard seed (Sin a 2), cashew nut (Ana o 2), hazelnut (Cor a 9), Brazil nut (Ber e 2), peanut (Ara h 3), and soybean (glycinin) (GenBank accession numbers AY846388, AF453947, AF449424, AY221641, AF093541, and X15122, respectively). (-) for gaps. Residues conserved in all proteins are in orange, and those conserved in at least 4 chains are in blue. The Asn cleavage site is indicated by an arrow and cysteines by asterisks. %I and %S, percentages of identity and similarity, respectively. Amino acid numbering for Sin a 2 is given for the mature protein (R1-V487).
Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci )
Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4. Fig 3
Comparison of peptides reported as epitopes in Ara h 330 with those corresponding to Sin a 2. Numerals indicate the position in each protein. Amino acids involved in the IgE binding in Ara h 3 epitopes are indicated by arrowheads. Conserved amino acids are shaded in black.
Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci )
Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5. Fig 4
Analysis of the expression and purification of rSin a 2 by SDS-PAGE. A, Time course of the production; immunoblotting with anti-polyhistidine antibody. B, Purified rSin a 2 was stained with Coomassie Bue (lane 1), and immunostained with the anti-polyhistidine antibody (lane 2) and with a pool of sera from patients with mustard allergy (lane 3). C, Purified nSin a 2 was stained with Coomassie Blue (lane 1) and immunostained with the same pool of sera (lane 2). Molecular mass markers are displayed in the left margin.
Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci )
Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6. Fig 5
Immunodetection of rSin a 2 and inhibition assays after SDS-PAGE and transference to membranes. A, IgE responses to rSin a 2 and nSin a 2 with sera from 8 patients with mustard allergy (lanes 1 to 8); C1 and C2 are responses to sera from a patient with olive pollen allergy and from a person without allergy, respectively. B, IgE immunodetection of purified rSin a 2 with a pool of sera from patients 4, 6, 7, and 8 after preincubation with 10 μg of BSA or with 10 μg of nSin a 2. C, Yellow mustard seed extract (20 μg total protein) was exposed to the same pool of sera preincubated with 10 μg of BSA or 10 μg of rSin a 2. Mustard seed extract (20 μg total protein) was stained with Coomassie Blue (CBS). Asterisks indicate the protein bands analyzed by MS and MS/MS. D, IgE immunodetection of nSin a 2 after preincubation of the individual sera with BSA (lane 1) or rSin a 2 (lane 2). Molecular mass markers are in the left margin.
Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci )
Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions